Apparatus and methods for sample acquisition

ABSTRACT

Apparatus and methods for sample acquisition, including for example, samples for flow cytometry systems. Certain embodiments include a plurality of plates, valves, and conduits. In particular embodiments, the plates are stacked and the conduits extend through stack of plates, and in specific embodiments each valve is in fluid communication with a conduit.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims the priority benefit of U.S. provisionalapplication No. 62/611,220, filed Dec. 28, 2017, the entire contents ofwhich is incorporated herein by reference.

FIELD OF THE INVENTION

Embodiments of the present invention relate to apparatus, methods andsystems for acquiring samples, including for example, samples forpolymerase chain reaction flow cytometry systems.

BACKGROUND

The following descriptions and examples are not admitted to be prior artby virtue of their inclusion within this section

Certain analysis systems, including for example polymerase chainreaction flow cytometry systems, can be operated to analyze individualsamples from a plurality of containers such as wells in a multiwellplate. In order to analyze samples from a large number of samplecontainers, each sample is typically removed from the sample containerand transported to the analysis system to be analyzed. In typicalexisting systems, this often requires a needle that moves from eachindividual sample container to aspirate the sample from one container.After aspirating a sample from one container, the needle then must bemoved to a second container to aspirate an additional sample.

The amount of time that is required to aspirate a large number ofsamples can be significant when an aspiration needle must be movedbetween each container to obtain the samples. This can result inincreased analysis times and affect the efficiency of such systems.Sample acquisition times can also be negatively affected by the need toclean or purge the system between samples. In addition, typical sampleacquisition systems can utilize complex mechanisms to move theaspiration needle to the different sample containers. Such mechanismscan involve numerous moving components that may be susceptible tomechanical failure. This can reduce the reliability of such sampleacquisition systems and result in unwanted delays in sample analysis.

While not limiting the scope of the present disclosure, variousembodiments of the present invention address issues of existing systems,including efficiency and reliability factors as noted above.

SUMMARY OF THE INVENTION

Exemplary embodiments of the present disclosure relate to methods,devices and systems for acquiring samples, including for example,samples for polymerase chain reaction flow cytometry systems.

Exemplary embodiments include an apparatus for sample acquisition. Incertain embodiments, the apparatus comprises: a stack of plates; aplurality of seals coupled to a first plate of the stack of plates,wherein each seal of the plurality of seals is configured to seal to awell in a well plate; a first plurality of conduits extending throughthe stack of plates and the plurality of seals; a plurality of gasvalves, wherein each valve is in fluid communication with a conduit inthe first plurality of conduits; a sample port in a second plate of thestack of plates; and a second plurality of conduits extending throughthe plurality of seals and the stack of plates, wherein the secondplurality of conduits is in fluid communication with the sample port.

In particular embodiments, the stack of plates comprises a first valveplate comprising a first plurality of check valves configured to directfluid in one direction in the second plurality of conduits. In someembodiments, the first plurality of check valves are arranged in aplurality of rows. In specific embodiments, the stack of platescomprises a second valve plate comprising a second plurality of checkvalves, and each check valve in the second plurality of check valves isin fluid communication with a row of the first plurality of checkvalves. In certain embodiments, the first plate is a first manifoldplate and the second plate is a valve housing plate. In particularembodiments, the stack of plates comprises a first valve plate and afirst channel plate, where the first valve plate is disposed between thefirst channel plate and the first manifold plate. In some embodiments,the stack of plates comprises a second manifold plate and a second valveplate, where the second manifold plate is disposed between the firstchannel plate and the second valve plate. In some embodiments, the stackof plates comprises a second channel plate, where the second channelplate is disposed between second valve plate and the valve housingplate.

In specific embodiments, the first manifold plate comprises: a firstside and a second side, where the second side is opposite the firstside; a first plurality of conduits configured to direct pressurized gasfrom the plurality of solenoid valves through the first manifold plate;and a second plurality of conduits between the first side and the secondside, wherein each conduit in the second plurality of conduits isarranged at an acute angle to the first side.

In certain embodiments, the first channel plate comprises a first sideand a second side, where the second side is opposite the first side; thefirst channel plate comprises a plurality of channels in the second sideof the first channel plate; the first valve plate comprises a pluralityof check valves arranged in a plurality of rows; each check valve in thefirst valve plate is aligned with a conduit of the second plurality ofconduits in the first manifold plate; and each row of check valves inthe first valve plate is aligned with a channel in the plurality ofchannels in the second side of the first channel plate. In particularembodiments, each check valve in the first valve plate is configured todirect fluid in one direction in a channel in the plurality of channelsin the second side of the first channel plate. In some embodiments, eachchannel in the plurality of channels in the second side of the firstchannel plate comprises an exit port that extends through the first sideof the first channel plate; and each check valve in a row of checkvalves of the first valve plate is configured to direct fluid toward theexit port of the channel to which the row of check valves is aligned.

In specific embodiments, the second manifold plate comprises a firstside and a second side, wherein the second side is opposite the firstside; the second manifold plate comprises a plurality of directionalports extending through the second manifold plate, where eachdirectional port is arranged at an acute angle to the first side of thesecond manifold plate; and the exit port of each channel in the firstchannel plate is aligned with a directional port in the second manifoldplate. In certain embodiments, the second valve plate comprises aplurality of check valves arranged in a row, and each directional portin the first channel plate is aligned with a check valve in the secondvalve plate.

In particular embodiments, the second channel plate comprises a firstside and a second side, where the second side is opposite the firstside; the second channel plate comprises an outlet channel in the secondside of the second channel plate, where the outlet channel is alignedwith the plurality of check valves of the second valve plate; the outletchannel of the channel valve plate comprises an outlet port; and eachcheck valve in the second valve plate is configured to direct fluidtoward the outlet port of the outlet channel.

In some embodiments, the outlet port of the second channel plate isaligned with a sample analysis port in the valve housing. In specificembodiments, the sample analysis port is in fluid communication with aflow cytometry analysis system. In certain embodiments, the plurality ofgas valves are configured as solenoid valves. Particular embodimentsfurther comprise a source of pressurized gas in fluid communication withthe plurality of gas valves. In some embodiments, the source ofpressurized gas comprises pressurized air. In specific embodiments, thesource of pressurized gas comprises pressurized nitrogen. In certainembodiments, the first valve plate and the second valve plate are formedfrom a flexible polymer. In particular embodiments, the first valveplate and the second valve plate are formed from polytetrafluoroethylene(PTFE). In some embodiments, the first channel plate and the secondchannel plate are formed from polytetrafluoroethylene (PTFE).

Specific embodiments further comprise a control system configured toopen and close the plurality of gas valves. In certain embodiments, theplurality of gas valves are arranged in a plurality of rows, the controlsystem is configured to sequentially open and close each gas valve in anindividual row of the plurality of rows. In particular embodiments, thecontrol system is configured to sequentially open and close each valvein a first row of the plurality of rows, and the control system isconfigured to sequentially open and close each valve in a second row ofthe plurality of rows after each valve in the first row has beensequentially opened and closed. In certain embodiments, the firstmanifold plate comprises a first side and a second side, wherein thesecond side is opposite the first side; the second side of the firstmanifold plate is engaged with well plate comprising a plurality ofwells, wherein each well comprises a sample; the plurality of gas valvesare in fluid communication with a source of pressurized gas; and thepressurized gas directs a portion of each sample from a well in theplurality of wells when a gas valve is open.

In particular embodiments, the portion of each sample is directed fromthe well in the plurality of wells, through the first manifold plate,the first valve plate, the first channel plate, the second manifoldplate, the second valve plate, the second channel plate and into thevalve housing. In some embodiments, the first manifold plate comprises afirst side and a second side, where the second side is opposite thefirst side; the first side of the first manifold plate is configured toengage the first valve plate; and the apparatus further comprises aplurality of sealing mechanisms coupled to the second side of the firstmanifold plate.

In certain embodiments, each sealing mechanism comprises an elastomerico-ring. In particular each sealing mechanism comprises a plug disposedwithin the elastomeric o-ring, and the plug comprises a first conduitand a second conduit extending through the plug. In some embodiments,each sealing mechanism in the plurality of sealing mechanisms comprises:a collar comprising a tapered inner surface; a plug comprising a lowersurface and a tapered outer surface, wherein the tapered outer surfaceof the plug is configured to engage the tapered inner surface of thecollar; a first conduit extending through the plug; and a second conduitextending through the plug. Specific embodiments further comprise a disccoupled to the lower surface of the plug, wherein the first conduit andthe second conduit extend through the disc.

In specific embodiments, each sealing mechanism in the plurality ofsealing mechanisms comprises: a collar comprising a tapered innersurface and a tapered outer surface; a plug comprising a lower surfaceand a tapered outer surface, wherein the tapered outer surface of theplug is configured to engage the tapered inner surface of the collar; adisc coupled to the lower surface of the plug; a first conduit extendingthrough the plug and the disc; and a second conduit extending throughthe plug and the disc. In certain embodiments, the disc and the taperedouter surface of the collar are configured to seal to a well in wellplate. Particular embodiments further comprise a plate comprising aplurality of magnets.

Certain embodiments include an apparatus for sample acquisition anddelivery, where the apparatus comprises: a first manifold plate, a firstvalve plate and a first channel plate, where the first valve plate isdisposed between the first channel plate and the first manifold plate; asecond manifold plate and a second valve plate, where the secondmanifold plate is disposed between the first channel plate and thesecond valve plate; a second channel plate and a valve housing, wherethe second channel plate is disposed between second valve plate and thevalve housing. Particular embodiments include a plurality of gas valvesdisposed in the valve housing, where each gas valve is configured toallow gas to flow through the valve housing, the second channel plate,the second valve plate, the second manifold plate, the first channelplate, the first valve plate and the first manifold plate when each gasvalve is in an open position.

In some embodiments, the first manifold plate comprises: a first sideand a second side, where the first side is opposite the second side; afirst plurality of conduits configured to direct pressurized gas fromthe plurality of solenoid valves through the first manifold plate; and asecond plurality of conduits between the second side and the first side,wherein each conduit in the second plurality of conduits is arranged atan acute angle to the first side. In specific embodiments, the firstchannel plate comprises a first side and a second side, wherein thesecond side is opposite the first side; the first channel platecomprises a plurality of channels in the second side of the firstchannel plate; the first valve plate comprises a plurality of checkvalves arranged in a plurality of rows; each check valve in the firstvalve plate is aligned with a conduit of the second plurality ofconduits in the first manifold plate; and each row of check valves inthe first valve plate is aligned with a channel in the plurality ofchannels in the second side of the first channel plate.

In certain embodiments, each check valve in the first valve plate isconfigured to direct fluid in one direction in a channel in theplurality of channels in the second side of the first channel plate. Inparticular embodiments, each channel in the plurality of channels in thesecond side of the first channel plate comprises an exit port thatextends through the first side of the first channel plate; and eachcheck valve in a row of check valves of the first valve plate isconfigured to direct fluid toward the exit port of the channel to whichthe row of check valves is aligned. In some embodiments, the secondmanifold plate comprises a first side and a second side, wherein thesecond side is opposite the first side; the second manifold platecomprises a plurality of directional ports extending through the secondmanifold plate, wherein each directional port is arranged at an acuteangle to the first side of the second manifold plate; and the exit portof each channel in the first channel plate is aligned with a directionalport in the second manifold plate. In specific embodiments, the secondvalve plate comprises a plurality of check valves arranged in a row; andeach directional port in the first channel plate is aligned with a checkvalve in the second valve plate.

In particular embodiments, the second channel plate comprises a firstside and a second side; the second channel plate comprises an outletchannel in the second side of the second channel plate, where the outletchannel is aligned with the plurality of check valves of the secondvalve plate; the outlet channel of the channel valve plate comprises anoutlet port; and each check valve in the second valve plate isconfigured to direct fluid toward the outlet port of the outlet channel.In some embodiments, the outlet port of the second channel plate isaligned with a sample analysis port in the valve housing. In specificembodiments, the sample analysis port is in fluid communication with aflow cytometry analysis system. In certain embodiments, the plurality ofgas valves are configured as solenoid valves. Particular embodiments,further comprise a source of pressurized gas in fluid communication withthe plurality of gas valves. In some embodiments, the source ofpressurized gas comprises pressurized air or pressurized nitrogen. Inspecific embodiments, the first valve plate and the second valve plateare formed from a flexible polymer.

In certain embodiments, the first valve plate and the second valve plateare formed from polytetrafluoroethylene (PTFE), and in particularembodiments the first channel plate and the second channel plate areformed from polytetrafluoroethylene (PTFE). Some embodiments furthercomprise a control system configured to open and close the plurality ofgas valves. In specific embodiments, the plurality of gas valves arearranged in a plurality of rows; and the control system is configured tosequentially open and close each gas valve in an individual row of theplurality of rows. In certain embodiments, the control system isconfigured to sequentially open and close each valve in a first row ofthe plurality of rows, and the control system is configured tosequentially open and close each valve in a second row of the pluralityof rows after each valve in the first row has been sequentially openedand closed.

In particular embodiments, the first manifold plate comprises a firstside and a second side; the second side of the first manifold plate isengaged with well plate comprising a plurality of wells, where each wellcomprises a sample; the plurality of gas valves are in fluidcommunication with a source of pressurized gas; and the pressurized gasdirects a portion of each sample from a well in the plurality of wellswhen a gas valve is open. In some embodiments, the portion of eachsample is directed from the well in the plurality of wells, through thefirst manifold plate, the first valve plate, the first channel plate,the second manifold plate, the second valve plate, the second channelplate and into the valve housing. In specific embodiments, the firstmanifold plate comprises a first side and a second side; the first sideof the first manifold plate is configured to engage the first valveplate; and the apparatus further comprises a plurality of sealingmechanisms coupled to the second side of the first manifold plate.

In certain embodiments, each sealing mechanism comprises an elastomerico-ring. In particular each sealing mechanism comprises a plug disposedwithin the elastomeric o-ring, and the plug comprises a first conduitand a second conduit extending through the plug. In some embodiments,each sealing mechanism in the plurality of sealing mechanisms comprises:a collar comprising a tapered inner surface; a plug comprising a lowersurface and a tapered outer surface, wherein the tapered outer surfaceof the plug is configured to engage the tapered inner surface of thecollar; a first conduit extending through the plug; and a second conduitextending through the plug. Specific embodiments further comprise a disccoupled to the lower surface of the plug, wherein the first conduit andthe second conduit extend through the disc.

In certain embodiments, each sealing mechanism in the plurality ofsealing mechanisms comprises: a collar comprising a tapered innersurface and a tapered outer surface; a plug comprising a lower surfaceand a tapered outer surface, wherein the tapered outer surface of theplug is configured to engage the tapered inner surface of the collar; adisc coupled to the lower surface of the plug; a first conduit extendingthrough the plug and the disc; and a second conduit extending throughthe plug and the disc. In particular embodiments, the disc and thetapered outer surface of the collar are configured to seal to a well inwell plate. Some embodiments further comprise a plate comprising aplurality of magnets.

Certain embodiments include a method of obtaining a plurality of samplesfor flow cytometry analysis, where the method comprises: positioning asample acquisition apparatus above a well plate comprising a pluralityof wells; and sequentially directing a flow of gas from the sampleacquisition apparatus into each well in the plurality of wells, wherethe flow of gas from the sample acquisition apparatus into each welldisplaces a portion of a sample from each well in the plurality of wellsinto the sample acquisition apparatus, and where the sample acquisitionapparatus does not move relative to the well plate. In particularembodiments, each portion of the sample from each well in the pluralityof wells directed into the sample acquisition apparatus is separated bya volume of gas from the flow of gas. In some embodiments, the sampleacquisition apparatus comprises: a plurality of plates; and a pluralityof valves configured to allow the flow of gas to pass through theplurality of plates, wherein each valve in the plurality of valves is influid communication with a well in the plurality of wells. In specificembodiments, each valve of the plurality of valves is initially in theclosed position; and sequentially directing the flow of gas from thesample acquisition apparatus into each well in the plurality of wellscomprises sequentially opening and closing the plurality of valves.

In certain embodiments, the plurality of valves are arranged in aplurality of rows, and sequentially opening and closing the plurality ofvalves comprises: opening and closing a first valve in a first row inthe plurality of valves; opening and closing a second valve in the firstrow of the plurality of valves after the first valve has been opened andclosed, wherein the second valve is adjacent to the first valve; andopening and closing each valve in the first row, wherein each valve isopened and closed after an adjacent valve has been opened and closed.Particular embodiments include: sequentially opening and closing theplurality of valves further comprises: opening and closing a first valvein a second row in the plurality of valves; opening and closing a secondvalve in the second row of the plurality of valves after the first valvehas been opened and closed, wherein the second valve is adjacent to thefirst valve; and opening and closing each valve in the second row,wherein each valve is opened and closed after an adjacent valve has beenopened and closed.

Specific embodiments include a flow cytometry system comprising: acuvette; a sample acquisition apparatus comprising a plurality of gasvalves, wherein the sample acquisition apparatus comprises a sample portin fluid communication with the cuvette; a gas manifold system in fluidcommunication with the plurality of gas valves; a control systemconfigured to sequentially open and close each gas valve in theplurality of gas valves; a sheath fluid supply system in fluidcommunication with the cuvette; and a drain manifold in fluidcommunication with the cuvette. In certain embodiments, the sample portis in fluid communication with the cuvette via a sample conduit, and theflow cytometry system further comprises a bubble detector configured todetect a bubble in the sample conduit. Particular embodiments furthercomprise a magnet configured to move relative to the sample conduit. Insome embodiments, the sheath fluid supply system and the drain manifoldare in fluid communication with a probe bath.

Specific embodiments include a flow cytometry system comprising: acuvette; a water supply; a sample proportional valve; a sheathproportional valve; a sample acquisition apparatus, wherein the sampleacquisition apparatus comprises a sample port in fluid communicationwith the cuvette, wherein the sample proportional valve is in fluidcommunication with the sample acquisition apparatus and the watersource; a sheath fluid supply system in fluid communication with thecuvette, wherein the sheath proportional valve is in fluid communicationwith the sheath fluid supply system and the water source; and a drainmanifold in fluid communication with the cuvette.

Certain embodiments include an apparatus for acquiring liquid samplesfrom a multiwell plate, the apparatus comprising: a first manifold platehaving a first side and a second side, wherein the second side isopposite the first side; a plurality of first conduits extending fromthe first side to the second side of the first manifold plate and aplurality of second conduits extending from the first side to the secondside of the first manifold plate, wherein the plurality of firstconduits and the plurality of second conduits are configured such thatwhen the first side of the first manifold plate is positioned adjacent amultiwell plate each well of the multiwell plate is in fluidcommunication with one of the plurality of first conduits and one of theplurality of second conduits; a plurality of gas valves in fluidcommunication with the plurality of first conduits; and a sample outletport in fluid communication with the plurality of second conduits. Inparticular embodiments, the multiwell plate is 96-well plate and theplurality of first conduits comprises 96 first conduits, the pluralityof second conduits comprises 96 second conduits, and the plurality ofgas valves comprises 96 gas valves. In some embodiments, the multiwellplate is 384-well plate and the plurality of first conduits comprises384 first conduits, the plurality of second conduits comprises 384second conduits, and the plurality of gas valves comprises 384 gasvalves. Specific embodiments, further comprise a pressurized gasreservoir, wherein the pressurized gas reservoir is in fluidcommunication with the plurality of gas valves and is selectively influid communication with the plurality of the plurality of firstconduits via the plurality of gas valves.

Certain embodiments further comprise a plurality of protrusions coupledto the first side of the first manifold plate and configured to insertat least partially into the wells of the multiwell plate to create asubstantially airtight seal between each well and the first and secondconduits in fluid communication with each well. In particularembodiments, the plurality of protrusions each comprise an obturatingring. In some embodiments, each of the first conduits are positioned ata first angle to the first manifold plate; and each of the secondconduits are positioned at a second angle to the first manifold plate.In specific embodiments, the first angle is approximately 90 degrees andthe second angle is approximately 45 degrees.

Certain embodiments include an air valve housing plate having a firstside and a second side, where the second side is opposite the firstside, and where the plurality of air valves are coupled to the firstside of the air valve housing plate; a third plurality of conduitsextending from the first side of the air valve housing plate to thesecond side of the air valve housing plate, wherein the plurality of airvalves are in fluid communication with the plurality of first conduitsin the sample manifold plate via the plurality of third conduits.Particular embodiments further comprise: a first channel plate; and asecond channel plate, where the first channel plate is disposed betweenthe first manifold plate and the plurality of gas valves, and where thesecond channel plate is disposed between the first channel plate and theplurality of gas valves. Specific embodiments further comprise: a firstvalve plate and a second valve plate, where: the first valve plate isdisposed between the first manifold plate and the first channel plate;and the second valve plate is disposed between the first channel plateand the second channel plate.

The term “coupled” is defined as connected, although not necessarilydirectly, and not necessarily mechanically. Two items are “coupleable”if they can be coupled to each other, and, when coupled, may still becharacterized as “coupleable.” Unless the context explicitly requiresotherwise, items that are coupleable are also decoupleable, andvice-versa. One non-limiting way in which a first structure iscoupleable to a second structure is for the first structure to beconfigured to be coupled (or configured to be coupleable) to the secondstructure.

The terms “a” and “an” are defined as one or more unless this disclosureexplicitly requires otherwise.

The term “substantially” and its variations (e.g., “approximately” and“about”) are defined as being largely but not necessarily wholly what isspecified (and include wholly what is specified) as understood by one ofordinary skill in the art. In any disclosed embodiment, the terms“substantially,” “approximately,” and “about” may be substituted with“within [a percentage] of” what is specified, where the percentageincludes 0.1, 1, 5, and 10 percent.

The terms “comprise” (and any form of comprise, such as “comprises” and“comprising”), “have” (and any form of have, such as “has” and“having”), “include” (and any form of include, such as “includes” and“including”) and “contain” (and any form of contain, such as “contains”and “containing”) are open-ended linking verbs. As a result, a method ordevice that “comprises,” “has,” “includes” or “contains” one or moresteps or elements possesses those one or more steps or elements, but isnot limited to possessing only those one or more elements. Likewise, astep of a method or an element of a device that “comprises,” “has,”“includes” or “contains” one or more features possesses those one ormore features, but is not limited to possessing only those one or morefeatures. For example, a system that comprises four channels may havemore than four channels.

Furthermore, a device or structure that is configured in a certain wayis configured in at least that way, but may also be configured in waysthat are not listed. Metric units may be derived from the English unitsprovided by applying a conversion and rounding to the nearestmillimeter.

The feature or features of one embodiment may be applied to otherembodiments, even though not described or illustrated, unless expresslyprohibited by this disclosure or the nature of the embodiments.

Any embodiment of any of the disclosed devices and methods can consistof or consist essentially of—rather thancomprise/include/contain/have—any of the described elements and/orfeatures and/or steps. Thus, in any of the claims, the term “consistingof” or “consisting essentially of” can be substituted for any of theopen-ended linking verbs recited above, in order to change the scope ofa given claim from what it would otherwise be using the open-endedlinking verb.

Other features and associated advantages will become apparent withreference to the following detailed description of specific embodimentsin connection with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

The following drawings illustrate by way of example and not limitation.For the sake of brevity and clarity, every feature of a given structuremay not be labeled in every figure in which that structure appears.Identical reference numbers do not necessarily indicate an identicalstructure. Rather, the same reference number may be used to indicate asimilar feature or a feature with similar functionality, as maynon-identical reference numbers.

FIG. 1 is a schematic of a schematic diagram for a flow cytometry systemcomprising an apparatus configured for sample acquisition and delivery.

FIG. 2 is a perspective view of an apparatus for acquiring samplesaccording to an exemplary embodiment.

FIG. 3 is an exploded view of the embodiment of FIG. 1.

FIG. 4 is a first section view of the embodiment of FIG. 1.

FIG. 5 is a second section view of the embodiment of FIG. 1.

FIG. 6 is a first section view of a first manifold plate of theembodiment of FIG. 1.

FIG. 7 is a second section view of a first manifold plate of theembodiment of FIG. 1.

FIG. 8 is a perspective view of a first valve plate of the embodiment ofFIG. 1.

FIG. 9 is a more detailed view of the first valve plate of FIG. 8.

FIG. 10 is a perspective view of a first channel plate of the embodimentof FIG. 1.

FIG. 11 is section view of a second manifold plate of the embodiment ofFIG. 1

FIG. 12 is a perspective view of a second valve plate of the embodimentof FIG. 1.

FIG. 13 is a section view of a second channel plate of the embodiment ofFIG. 1

FIG. 14 is a first section view of first valve housing plate of theembodiment of FIG. 1.

FIG. 15 is a second section view of first valve housing plate of theembodiment of FIG. 1

FIG. 16 is a section view of second valve housing plate of theembodiment of FIG. 1.

FIG. 17 is a detailed view of the first valve plate of FIGS. 8 and 9during operation.

FIG. 18 is a section view of a sealing mechanism of the embodiment ofFIG. 1.

DETAILED DESCRIPTION

Various features and advantageous details are explained more fully withreference to the non-limiting embodiments that are illustrated in theaccompanying drawings and detailed in the following description. Itshould be understood, however, that the detailed description and thespecific examples, while indicating embodiments of the invention, aregiven by way of illustration only, and not by way of limitation. Varioussubstitutions, modifications, additions, and/or rearrangements willbecome apparent to those of ordinary skill in the art from thisdisclosure.

In the following description, numerous specific details are provided toprovide a thorough understanding of the disclosed embodiments. One ofordinary skill in the relevant art will recognize, however, that theinvention may be practiced without one or more of the specific details,or with other methods, components, materials, and so forth. In otherinstances, well-known structures, materials, or operations are not shownor described in detail to avoid obscuring aspects of the invention. Itis understood that for purposes of clarity, not all reference numbersare shown for every component visible in each figure.

It should be understood that the present devices and methods are notintended to be limited to the particular forms disclosed. Rather, theyare to cover all modifications, equivalents, and alternatives fallingwithin the scope of the claims.

Referring initially to FIG. 1, a schematic diagram for a flow cytometrysystem 50 comprises an apparatus 100 configured for sample acquisitionand delivery.

An overview of the operation of apparatus 100 will be presentedinitially, followed by further discussion of individual components. Ingeneral, apparatus 100 can rapidly acquire samples from a plurality ofreservoirs, including for example, wells in a well plate. As usedherein, the term “sample” or “sample fluid” is intended to beinterpreted broadly to include a portion (including all) of a volume ofmatter comprising liquid contained in a reservoir. Apparatus 100 can beincorporated in a flow cytometry system to provide for rapid sampleanalysis.

Apparatus 100 includes a stack of plates that can be coupled to a wellplate and individually seal each well in the well plate. As used herein,the terms “plate” and “plates” is intended to be construed broadly, andrefers to any generally planar structure without limiting to aparticular shape, thickness, or rigidity. For example, certaincomponents in the disclosed apparatus may be referred to as “valveplates”, which may be formed from sheets of flexible (e.g. polymeric)material.

Apparatus 100 also includes a first set of conduits that extend from aset of valves, through the plates and into each well. Each valve iscoupled to a pressurized gas supply, so that as each valve is opened,pressurized gas is directed into a well. In particular embodiments, thevalves may be solenoid valves. The pressurized gas directs fluid fromeach well into a second set of conduits that extend through the stack ofplates and to a sample port for analysis by the flow cytometry system.The valves can be sequentially opened and closed to provide efficientsample acquisition and delivery for analysis. Exemplary embodiments arenot limited to the orientation shown in the figures. For ease ofunderstanding the operation of apparatus 100, with respect to theincluded figures the gas is directed downward through the apparatus,while the sample fluid is directed upward from the sample container(e.g. well) to the sample port. It is understood the “downward” and“upward” description is only for purpose of explanation with respect tothe attached figures and is not intended to limit exemplary embodimentsto any particular orientation.

Unlike typical existing systems, apparatus 100 does not include apipette or other component that must be indexed to individual wells toaspirate a sample. Apparatus 100 therefore includes fewer componentsthat move relative to a well plate from which samples are acquired. Asexplained in more detail below, apparatus 100 comprises angled conduitsand thin plates of flexible material that include check valves to directsample flow from the wells to the sample port.

Referring now to FIGS. 2-5, apparatus 100 includes a stacked assembly ofplates comprising (from bottom to top) a first manifold plate 110, afirst valve plate 120, a first channel plate 130, a second manifoldplate 140, a second valve plate 150, a second channel plate 160, and avalve housing 170. In this embodiment, valve housing 170 comprises afirst valve housing plate 180 and a second valve housing plate 190. Forpurposes of clarity, certain components of apparatus 100 are not shownor labeled in each in the assembly and exploded views, but included inthe section views discussed further below.

During operation of apparatus 100, pressurized gas (e.g. air ornitrogen) is directed to a gas valve 200 that can be controlled to openand allow the gas to flow through conduits 219 into a well in a wellplate. For purposes of clarity, only a single gas valve 200 is shown inFIGS. 2 and 4. It is understood that each aperture 195 in second valvehousing plate 190 may comprise a gas valve 200. In the embodiment shown,second valve housing plate 190 comprises apertures 195 in an arrangementof twelve rows 201-212 of eight apertures each that are configured tocorrespond with a 96 well plate. It is understood that other embodimentsmay comprise other arrangements, including for example, a 384 wellplate.

In the embodiment shown, each conduit 219 comprises a plurality ofapertures or conduits in each of the plates in apparatus 100 andincludes a tube 235 that extends into each sample container (e.g. well).In the illustrated embodiment, each conduit 219 comprises conduit 119 infirst manifold plate 110 (shown in FIG. 6), aperture 129 in first valveplate 120 (FIG. 9), aperture 139 in first channel plate 130 (FIG. 10),aperture 149 in second manifold plate 140 (FIG. 11), aperture 159 insecond valve plate 150 (FIG. 12), aperture 169 in second channel plate160 (FIG. 13), and conduit 189 in first valve housing plate 180 (FIG.15). As shown in FIG. 15, first air valve housing plate 180 comprisesapertures 185 extending between a first side 181 and a second side 182that is opposite of first side 181. Air valves 200 can extend throughsecond air valve housing plate 170 and be coupled to first side 181 offirst valve housing plate 180. Air valves 200 are in fluid communicationwith conduits 119 in first manifold plate 110 via conduits 189 in firstvalve housing plate 180 (and additional conduits and apertures in theplates as described above).

The pressurized gas from each gas valve 200 is directed through aconduit 213 and forces the sample fluid from the well into an angledconduit 118 in first manifold plate (also referred to as a samplemanifold plate) 110 as shown in FIG. 7. First manifold plate comprises afirst side 116 opposite a second side 117, and each of the plurality ofangled conduits 118 is arranged at an acute angle to first side 116. Thesample fluid exits angled conduit 118 and passes through valves 128 infirst valve plate 120 shown in FIGS. 8 and 9 (a more detailed view ofvalve 128 during operation is shown in FIG. 17). As shown in FIG. 17,valve 128 is lifted by the sample fluid flow (indicated by arrow 101)such that valve 128 seals against first channel plate 130. In theconfiguration shown, first valve plate 120 is located between firstmanifold plate 110 and first channel plate 130.

After passing through valves 128 in first valve plate 120, the samplefluid is directed to channels 138 formed in a second side 132 of firstchannel plate 130 shown in FIG. 10. Channels 138 do not extendcompletely through first channel plate 130, but channels 138 are influid communication with an exit port or conduit 136 that extends fromsecond side 132 of channel plate 130 to a first side 131 of firstchannel plate 130 that is opposite of second side 132. When apparatus100 is assembled, angled conduits 118 in first manifold plate 110 areangled toward conduits 136 shown in FIG. 10. This angled configuration(along with the flap configuration of valve 128) allows sample fluidflow to pass through valves 128 and toward conduits 136 without backflowing through channel 138 (e.g. flowing away from conduits 136).

After exiting conduits 136 from first channel plate 130, the samplefluid is directed through directional ports or conduits 148 throughsecond manifold plate 140 shown in FIG. 11. Second manifold platecomprises a first side 146 opposite a second side 147, and each of theplurality of angled conduits 148 is arranged at an acute angle to firstside 146. Similar to angled conduits 118, conduits 148 are also angledto direct the sample fluid in the desired direction. From conduits 148,the sample fluid then passes through valves 158 in second valve plate150 shown in FIG. 12. Valves 158 are configured similar to valves 128and allow the sample fluid to pass into an outlet or cross flow channel168 in second channel plate 160 in the desired direction (e.g. toward anoutlet port or conduit 166). Cross flow channel 168 is in fluidcommunication with conduit 166 that extends from a second side 162 ofsecond channel plate 160 to a first side 161 of second channel plate160. Conduit 166 is in further fluid communication with a sample port178 in first valve housing plate 180 of valve housing 170. In certainembodiments, sample port 178 can be accessed by a flow cytometry systemfor analysis of the sample fluid.

Referring now to FIG. 18, exemplary embodiments may comprise a sealingmechanism 215 configured to seal apparatus 100 (in particular firstmanifold plate 110) to a sample container, including for example, a wellin a well plate. In the embodiment shown, sealing mechanism 215comprises a collar or obturating ring 211 surrounding a plug or centralportion 212. Sealing mechanism 215 can further comprise a disc 216 thatcan be coupled to first manifold plate 110 (as shown in FIG. 4). Otherembodiments may comprise a sealing mechanism configured as a gasket withopenings corresponding to wells in a well plate.

In specific embodiments, obturating ring may be formed from a softermaterial than central portion 112. Central portion 212 can comprise aconduit 213 configured to allow pressurized gas to pass into a samplecontainer (e.g. a sample well). Central portion 212 can also comprise aconduit 214 configured to allow sample fluid to pass from the samplecontainer to angled conduit 118 in first manifold plate 110. Ring 211can comprise a tapered inner surface 217 and a tapered outer surface218, while central portion 212 comprises a tapered outer surface 219configured to engage tapered inner surface 217 of ring 211. Duringoperation of apparatus 100, sealing mechanism 215 can effectively sealapparatus 100 to allow pressurized gas to direct sample fluid from asample container and into apparatus 100.

Exemplary embodiments of the present disclosure provide significantbenefits over typical sample acquisition apparatus and methods. Forexample, exemplary embodiments can reduce the sample acquisition time,which can reduce the time and costs associated with sample preparationprocesses. This is particularly true of processes that include a highnumber of cycles, including for example, flow cytometry processes.

In particular embodiments, apparatus 100 can be operated such that eachvalve 200 in a particular row of valves is sequentially opened andclosed prior to operating valves from any other rows. In a specificembodiment, each valve 200 in a first row is sequentially opened,followed by each valve 200 in a second row being sequentially opened,until each valve 200 in each row has been sequentially opened. Byfollowing a known pattern of valve operation, the order of the acquiredsamples can also be determined.

In some embodiments, each gas valve 200 may be opened for a period oftime sufficient to evacuate the entire sample from the sample containerand to introduce a gas bubble into angled conduit 118 (and thesubsequent conduits and channels through which the sample fluid flows).Accordingly, a gas bubble can separate each sample evacuated from aparticular sample container. In specific embodiments, flow cytometrysystem 50 may comprise a bubble detector 51 to detect a bubble betweendifferent samples. Apparatus 100 can therefore allow samples frommultiple sample containers to be distinguished quickly and accuratelywithout having to move an aspirating needle to each sample container toremove the sample.

The above specification and examples provide a complete description ofthe structure and use of an exemplary embodiment. Although certainembodiments have been described above with a certain degree ofparticularity, or with reference to one or more individual embodiments,those skilled in the art could make numerous alterations to thedisclosed embodiments without departing from the scope of thisinvention. As such, the illustrative embodiment of the present devicesis not intended to be limited to the particular forms disclosed. Rather,they include all modifications and alternatives falling within the scopeof the claims, and embodiments other than the one shown may include someor all of the features of the depicted embodiment. Further, whereappropriate, aspects of any of the examples described above may becombined with aspects of any of the other examples described to formfurther examples having comparable or different properties andaddressing the same or different problems. Similarly, it will beunderstood that the benefits and advantages described above may relateto one embodiment or may relate to several embodiments.

The claims are not to be interpreted as including means-plus- orstep-plus-function limitations, unless such a limitation is explicitlyrecited in a given claim using the phrase(s) “means for” or “step for,”respectively.

REFERENCES

The following references are incorporated herein by reference:

-   U.S. Pat. No. 5,560,811-   U.S. Pat. No. 6,042,709-   U.S. Pat. No. 6,149,787-   U.S. Pat. No. 7,024,281-   U.S. Pat. Pub. 2004/0071602-   U.S. Pat. Pub. 2005/0238545-   U.S. Pat. Pub. 2006/0198765-   U.S. Pat. Pub. 2017/0199210

We claim:
 1. An apparatus for sample acquisition, the apparatuscomprising: a stack of plates; a plurality of seals coupled to a firstplate of the stack of plates, wherein each seal of the plurality ofseals is configured to seal to a well in a well plate; a first pluralityof conduits extending through the stack of plates and the plurality ofseals; a plurality of gas valves, wherein each valve is in fluidcommunication with a conduit in the first plurality of conduits; asample port in a second plate of the stack of plates; and a secondplurality of conduits extending through the plurality of seals and thestack of plates, wherein the second plurality of conduits is in fluidcommunication with the sample port.
 2. The apparatus of claim 1 whereinthe stack of plates comprises a first valve plate comprising a firstplurality of check valves configured to direct fluid in one direction inthe second plurality of conduits.
 3. The apparatus of claim 2 whereinthe first plurality of check valves are arranged in a plurality of rows.4. The apparatus of claim 3 wherein: the stack of plates comprises asecond valve plate comprising a second plurality of check valves; andeach check valve in the second plurality of check valves is in fluidcommunication with a row of the first plurality of check valves.
 5. Theapparatus of claim 1 wherein: the first plate is a first manifold plateand the second plate is a valve housing plate, and wherein the stack ofplates comprises: a first valve plate; a first channel plate, whereinthe first valve plate is disposed between the first channel plate andthe first manifold plate; a second manifold plate; a second valve plate,wherein the second manifold plate is disposed between the first channelplate and the second valve plate; and a second channel plate, whereinthe second channel plate is disposed between second valve plate and thevalve housing plate.
 6. The apparatus of claim 5 wherein the firstmanifold plate comprises: a first side and a second side, wherein thesecond side is opposite the first side; a first plurality of conduitsconfigured to direct pressurized gas from the plurality of solenoidvalves through the first manifold plate; and a second plurality ofconduits between the first side and the second side, wherein: eachconduit in the second plurality of conduits is arranged at an acuteangle to the first side; the first channel plate comprises a first sideand a second side, wherein the second side is opposite the first side;the first channel plate comprises a plurality of channels in the secondside of the first channel plate; the first valve plate comprises aplurality of check valves arranged in a plurality of rows; each checkvalve in the first valve plate is aligned with a conduit of the secondplurality of conduits in the first manifold plate; and each row of checkvalves in the first valve plate is aligned with a channel in theplurality of channels in the second side of the first channel plate. 7.The apparatus of claim 6 wherein: each check valve in the first valveplate is configured to direct fluid in one direction in a channel in theplurality of channels in the second side of the first channel plate;each channel in the plurality of channels in the second side of thefirst channel plate comprises an exit port that extends through thefirst side of the first channel plate; each check valve in a row ofcheck valves of the first valve plate is configured to direct fluidtoward the exit port of the channel to which the row of check valves isaligned; the second manifold plate comprises a first side and a secondside, wherein the second side is opposite the first side; the secondmanifold plate comprises a plurality of directional ports extendingthrough the second manifold plate, wherein each directional port isarranged at an acute angle to the first side of the second manifoldplate; the exit port of each channel in the first channel plate isaligned with a directional port in the second manifold plate; the secondvalve plate comprises a plurality of check valves arranged in a row; andeach directional port in the first channel plate is aligned with a checkvalve in the second valve plate.
 8. The apparatus of claim 7 wherein:the second channel plate comprises a first side and a second side,wherein the second side is opposite the first side; the second channelplate comprises an outlet channel in the second side of the secondchannel plate, wherein the outlet channel is aligned with the pluralityof check valves of the second valve plate; the outlet channel of thechannel valve plate comprises an outlet port; each check valve in thesecond valve plate is configured to direct fluid toward the outlet portof the outlet channel; and the outlet port of the second channel plateis aligned with a sample analysis port in the valve housing.
 9. Theapparatus of claim 8 wherein the sample analysis port is in fluidcommunication with a flow cytometry analysis system.
 10. The apparatusof claim 5 wherein the plurality of gas valves are configured assolenoid valves.
 11. The apparatus of claim 5 further comprising asource of pressurized gas in fluid communication with the plurality ofgas valves.
 12. The apparatus of claim 5 further comprising a controlsystem configured to open and close the plurality of gas valves,wherein: the plurality of gas valves are arranged in a plurality ofrows; the control system is configured to sequentially open and closeeach gas valve in an individual row of the plurality of rows; thecontrol system is configured to sequentially open and close each valvein a first row of the plurality of rows; the control system isconfigured to sequentially open and close each valve in a second row ofthe plurality of rows after each valve in the first row has beensequentially opened and closed; the first manifold plate comprises afirst side and a second side, wherein the second side is opposite thefirst side; the second side of the first manifold plate is engaged withwell plate comprising a plurality of wells, wherein each well comprisesa sample; the plurality of gas valves are in fluid communication with asource of pressurized gas; the pressurized gas directs a portion of eachsample from a well in the plurality of wells when a gas valve is open;and the portion of each sample is directed from the well in theplurality of wells, through the first manifold plate, the first valveplate, the first channel plate, the second manifold plate, the secondvalve plate, the second channel plate and into the valve housing. 13.The apparatus of claim 5 wherein: the first manifold plate comprises afirst side and a second side, wherein the second side is opposite thefirst side; the first side of the first manifold plate is configured toengage the first valve plate; and the apparatus further comprises aplurality of sealing mechanisms coupled to the second side of the firstmanifold plate.
 14. The apparatus of claim 13 wherein each sealingmechanism in the plurality of sealing mechanisms comprises: a collarcomprising a tapered inner surface; a plug comprising a lower surfaceand a tapered outer surface, wherein the tapered outer surface of theplug is configured to engage the tapered inner surface of the collar; afirst conduit extending through the plug; a second conduit extendingthrough the plug; and a disc coupled to the lower surface of the plug,wherein the first conduit and the second conduit extend through thedisc.
 15. The apparatus of claim 13 wherein each sealing mechanism inthe plurality of sealing mechanisms comprises: a collar comprising atapered inner surface and a tapered outer surface; a plug comprising alower surface and a tapered outer surface, wherein the tapered outersurface of the plug is configured to engage the tapered inner surface ofthe collar; a disc coupled to the lower surface of the plug; a firstconduit extending through the plug and the disc; and a second conduitextending through the plug and the disc.
 16. The apparatus of claim 15wherein the disc and the tapered outer surface of the collar areconfigured to seal to a well in well plate.
 17. The apparatus of claim 5further comprising a plate comprising a plurality of magnets.